The BioCD is an emerging label-free assay technology with potential for high multiplexing and high throughput to screen for many analytes across many samples simultaneously. The basis of the BioCD technology is rapid optical interferometric scanning. Interferometry is the most sensitive and quantitative means of direct optical detection. It is faster than fluorescence, with better signal-to-noise, and it requires no labels, which is essential for multiple analyte detection. The broad, long-term objectives of this proposal are to apply the BioCD for the first time to the prognosis of cancer. The goal is to assay multiple biomarkers across a large cohort of 300 patients, at a level previously inaccessible to immunohistological arrays, to predict patient outcome in response to chemotherapy. The specific aims of this proposal are to establish standard response curves for a set of biomarkers relevant for predicting chemotherapy outcome for patients with acute lymphocytic leukemia (ALL), and to do a comparative study between assays performed on the BioCD and assays performed previously on a limited number of tissue microarrays. This is followed by the aim to scale up the capacity of the BioCD to screen for an expanded biomarker set across a set of samples already collected from the cohort of 300 patients with acute lymphocytic leukemia. The research design and the methods for achieving the stated goals rely on high- capacity automated protein spotters and high-speed laser interferometric readers. Standard concentration curves will be established using commercially available antigens in tissue lysates, followed by measurements of marker concentrations in healthy tissue lysate. These measurements set the gold standard against which the expanded marker set measured across the large patient cohort will be compared. The relevance of this research to public health is the expansion of the marker and sample base to establish stronger clinical confirmation of the prognostic value of inactivation of a molecular pathway in patients with standard-risk acute lymphocytic leukemia, and to establish the BioCD as a novel high-capacity resource for diagnostic and prognostic applications for cancer.